MOZ/ENL complex is a recruiting factor of leukemic AF10 fusion proteins

Changes in the transcriptional machinery cause aberrant self-renewal of non-stem hematopoietic progenitors. AF10 fusions, such as CALM-AF10, are generated via chromosomal translocations, causing malignant leukemia. In this study, we demonstrate that AF10 fusion proteins cause aberrant self-renewal via ENL, which binds to MOZ/MORF lysine acetyltransferases (KATs). The interaction of ENL with MOZ, via its YEATS domain, is critical for CALM-AF10-mediated leukemic transformation. The MOZ/ENL complex recruits DOT1L/AF10 fusion complexes and maintains their chromatin retention via KAT activity. Therefore, inhibitors of MOZ/MORF KATs directly suppress the functions of AF10 fusion proteins, thereby exhibiting strong antitumor effects on AF10 translocation-induced leukemia. Combinatorial inhibition of MOZ/MORF and DOT1L cooperatively induces differentiation of CALM-AF10-leukemia cells. These results reveal roles for the MOZ/ENL complex as an essential recruiting factor of the AF10 fusion/DOT1L complex, providing a rationale for using MOZ/MORF KAT inhibitors in AF10 translocation-induced leukemia.

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All of the IP -western, ChIP-qPCR, RT-qPCR, and gRNA competition assay experiments were performed at least twice and confirmed their reproducibility. Myeloid progenitor transformation assay, and drug sensitivity analysis were performed at least three times (>3 biological replicates). RNA-seq analysis was performed for three biological replicates for each sample type. ChIP-seq analysis was performed once for most of the samples, and its reproducibility was confirmed by ChIP-qPCR analysis on selected targets. ChIP-seq analysis for ENL in HEK293T was performed twice for reproducibility. The other experiments were performed at least twice.
Randomization was not applied to the in vivo drug sensitivity tests, as the mice carrying leukemia cells were evenly distributed to each experimental group based on the luminescent signals before the drug treatment.
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Female C57BL/6JJcl mice were obtained from CLEA Japan inc.. Five-week-old female C57BL/6JJcl mice were used for bone marrow extraction and seven to eight-week-old mice were used for leukemogenesis assay. Mice were allowed free access to food and water and were maintained at room temperature (about 25C) with constant humidity (about 50%) on a 12-hours light/dark cycle.
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March 2021
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